title: Characterization of alpha-fetoprotein in fetal striped dolphin (Stenella coeruleoalba): purification of protein product and molecular cloning of the corresponding transcript.
abstract: Alpha-fetoprotein (AFP) is a fetal glycoprotein that is known as a biomarker for monitoring pregnancy in many mammalian species. However, characterization of AFP has not yet been undertaken in any cetacean species. Here, we purified AFP from the serum of fetal striped dolphin by chemical precipitation followed by a combination of immunoadsorbent column chromatography and gel filtration. The molecular masses of native and denatured dolphin AFP were estimated to be ∼78,000 Da by gel filtration and ∼68,000 Da by SDS-PAGE, respectively, representing typical masses reported for mammalian AFPs. In fetal serum, only the AFP band (∼68,000 Da) appeared to be immunoreactive to an antiserum against purified dolphin AFP, indicating sufficient specificity for the development of an AFP immunoassay. Full-length cDNA encoding for the dolphin AFP was cloned from fetal liver and revealed an open reading frame comprising 610 amino acid residues, which included a putative signal peptide of 18 amino acid residues. This was followed by a sequence identical to the N-terminus of purified AFP. The deduced amino acid sequence of dolphin AFP showed more than 80% identity to those of other mammalian AFPs. To our knowledge, the present report represents the first identification and characterization of AFP from any cetacean species.
authors.affiliation: Graduate School of Fisheries Sciences, Hokkaido University, Hakodate, Hokkaido 041-8611, Japan.
authors.forename: Yuka
authors.initials: Y
authors.lastname: Morita
authors.forename: Naoshi
authors.initials: N
authors.lastname: Hiramatsu
authors.forename: Toshiaki
authors.initials: T
authors.lastname: Fujita
authors.forename: Haruna
authors.initials: H
authors.lastname: Amano
authors.forename: Takashi
authors.initials: T
authors.lastname: Todo
authors.forename: Akihiko
authors.initials: A
authors.lastname: Hara
mesh_terms: D000220:Adaptation, Biological; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004289:Dolphins / Q000196:embryology* / Q000378:metabolism; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D007106:Immune Sera; D008099:Liver / Q000378:metabolism; D008969:Molecular Sequence Data; D010802:Phylogeny; D010920:Placenta / Q000378:metabolism; D011247:Pregnancy; D013045:Species Specificity; D000509:alpha-Fetoproteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism
chemical_list: D007106:Immune Sera; D000509:alpha-Fetoproteins